Structural analysis of specific metal chelating inhibitor binding to the endonuclease domain of influenza pH1N1 (2009) polymerase.
Structural analysis of specific metal chelating inhibitor binding to the endonuclease domain of influenza pH1N1 (2009) polymerase.
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DOI：
10.1371/journal.ppat.1002831
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发表时间：
2012
期刊：
影响因子：
6.7
通讯作者：
Cusack S
中科院分区：
文献类型：
--
作者：
Kowalinski E;Zubieta C;Wolkerstorfer A;Szolar OH;Ruigrok RW;Cusack S
人们普遍认为，新的抗病毒药物，不易产生耐药性，将是一个理想的替代目前的药物选择，以便能够治疗潜在的严重流感感染。进行RNA基因组转录和复制的病毒聚合酶是抗病毒药物的一个有吸引力的靶点，因为有效的聚合酶抑制剂可以在早期直接阻止病毒复制。最近对异源三聚体聚合酶的功能结构域的结构研究，其包括亚基PA、PB1和PB2，为基于结构的方法优化病毒复制抑制剂开辟了道路。特别地，病毒转录的独特的夺帽机制可以通过靶向PB2帽结合结构域或PA核酸内切酶结构域来抑制。在这里，我们描述了2009年大流行的H1N1（pH1N1）PA核酸内切酶结构域与一系列特异性抑制剂，包括四个二酮化合物和一个绿色茶儿茶素，所有这些螯合酶的活性位点的两个关键锰离子的高分辨率X射线共晶体结构。不同化合物的结合模式和磷酸单甘肽的结合模式的比较突出了，首先，碱性金属结合支架上的不同取代基可以如何定向以结合在活性位点空腔内的不同子口袋中，其次，活性位点空腔的某些结构元件的可塑性，这导致诱导的适合结合。这些结果对于优化针对流感病毒聚合酶的夺帽核酸内切酶活性的更有效的抑制剂的设计将是重要的。2009年流感大流行、高致病性H5N1禽流感病毒株的持续潜在威胁以及对靶向神经氨酸酶或M2离子通道的当前抗流感药物的耐药性的广泛发生，都突出了在缺乏疫苗接种保护的情况下治疗严重流感感染的替代治疗选择的需要。进行RNA基因组转录和复制的病毒聚合酶是新型抗病毒药物的有吸引力的靶标，因为有效的聚合酶抑制剂将直接阻止复制。异源三聚体聚合酶通过独特的夺帽机制进行转录，该机制涉及分别由PB2和PA亚基进行的宿主前mRNA帽结合和内切核酸裂解。PB2帽结合和PA核酸酶结构域的晶体结构现在都是可用的，允许帽抢夺抑制剂的结构指导优化。在这里，我们提出了一系列的共晶体结构的2009年大流行H1N1 PA核酸内切酶结构域，揭示了几个已知的核酸内切酶抑制剂的结合模式。所有抑制剂螯合核酸酶的活性位点中的两个锰离子，但金属结合支架的不同延伸结合在活性位点空腔的不同子口袋中。这些结果突出了基于结构的方法对开发更有效的流感聚合酶抑制剂的价值。
It is generally recognised that novel antiviral drugs, less prone to resistance, would be a desirable alternative to current drug options in order to be able to treat potentially serious influenza infections. The viral polymerase, which performs transcription and replication of the RNA genome, is an attractive target for antiviral drugs since potent polymerase inhibitors could directly stop viral replication at an early stage. Recent structural studies on functional domains of the heterotrimeric polymerase, which comprises subunits PA, PB1 and PB2, open the way to a structure based approach to optimise inhibitors of viral replication. In particular, the unique cap-snatching mechanism of viral transcription can be inhibited by targeting either the PB2 cap-binding or PA endonuclease domains. Here we describe high resolution X-ray co-crystal structures of the 2009 pandemic H1N1 (pH1N1) PA endonuclease domain with a series of specific inhibitors, including four diketo compounds and a green tea catechin, all of which chelate the two critical manganese ions in the active site of the enzyme. Comparison of the binding mode of the different compounds and that of a mononucleotide phosphate highlights, firstly, how different substituent groups on the basic metal binding scaffold can be orientated to bind in distinct sub-pockets within the active site cavity, and secondly, the plasticity of certain structural elements of the active site cavity, which result in induced fit binding. These results will be important in optimising the design of more potent inhibitors targeting the cap-snatching endonuclease activity of influenza virus polymerase. The 2009 influenza pandemic, the on-going potential threat of highly pathogenic H5N1 avian strains and the widespread occurrence of resistance to current anti-influenza drugs targeting the neuraminidase or the M2 ion channel, all highlight the need for alternative therapeutic options to treat serious influenza infections in the absence of protection by vaccination. The viral polymerase, which performs transcription and replication of the RNA genome, is an attractive target for novel antiviral drugs since potent polymerase inhibitors will directly stall replication. The heterotrimeric polymerase performs transcription by a unique cap-snatching mechanism, which involves host pre-mRNA cap-binding and endonucleolytic cleavage by the PB2 and PA subunits respectively. Crystal structures of both the PB2 cap-binding and PA nuclease domains are now available allowing structure-guided optimisation of cap-snatching inhibitors. Here we present a series of co-crystal structures of the 2009 pandemic H1N1 PA endonuclease domain that reveal the binding mode of several known endonuclease inhibitors. All inhibitors chelate the two manganese ions in the active site of the nuclease but different extensions to the metal binding scaffold bind in distinct sub-pockets of the active site cavity. These results highlight the value of structure-based approaches to the development of more potent influenza polymerase inhibitors.
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影响因子：
3.7
作者：
Iwai Y;Murakami K;Gomi Y;Hashimoto T;Asakawa Y;Okuno Y;Ishikawa T;Hatakeyama D;Echigo N;Kuzuhara T
通讯作者：
Kuzuhara T
影响因子：
4.9
作者：
Hastings, JC;Selnick, H;Tomassini, JE
通讯作者：
Tomassini, JE
影响因子：
5.8
作者：
Gouet, P;Courcelle, E;Métoz, F
通讯作者：
Métoz, F
DOI：
10.1073/pnas.1121297109
发表时间：
2012-02-14
影响因子：
11.1
作者：
Palese, Peter;Wang, Taia T.
通讯作者：
Wang, Taia T.
影响因子：
4.8
作者：
Klumpp, K;Doan, L;Handa, B
通讯作者：
Handa, B